Generation of induced pluripotent stem cells from muscular dystrophy patients: efficient integration-free reprogramming of urine derived cells. J Vis Exp 2015 Jan 28(95):52032
Date
02/05/2015Pubmed ID
25650629Pubmed Central ID
PMC4354557DOI
10.3791/52032Scopus ID
2-s2.0-84923595731 (requires institutional sign-in at Scopus site) 34 CitationsAbstract
Dystrophic cardiomyopathy is a poorly understood consequence of muscular dystrophy. Generating induced Pluripotent Stem Cells (iPSCs) from patients with muscular dystrophy is an invaluable cellular source for in vitro disease model systems and can be used for drug screening studies. Patient-derived urine cells have been used in successful reprogramming into induced pluripotent stem cells in order to model dystrophic cardiomyopathy(1). Addressing the safety concerns of integrating vector systems, we present a protocol using a non-integrating Sendai virus vector for transduction of Yamanaka factors into urine cells collected from patients with muscular dystrophy. This protocol generates fully reprogrammed clones within 2-3 weeks. The pluripotent cells are vector-free by passage-13. These dystrophic iPSCs can be differentiated into cardiomyocytes and used either to study disease mechanisms or for drug screening.
Author List
Afzal MZ, Strande JLMESH terms used to index this publication - Major topics in bold
Cell DifferentiationCellular Reprogramming
Cellular Reprogramming Techniques
Genetic Vectors
Humans
Induced Pluripotent Stem Cells
Muscular Dystrophies
Myocytes, Cardiac
Sendai virus
Transduction, Genetic
